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A kind of method and kit for detecting dha content in blood sample

A technology for blood samples and detection samples, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of operator hazards, experimental waste liquid discharge does not conform to the concept of environmental protection, etc., achieve rapid and accurate analysis, easy clinical promotion and popularization, High throughput effect

Active Publication Date: 2019-01-29
JINAN YING SHENG BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As we all know, acetonitrile is a highly toxic compound, frequent contact with acetonitrile is a great hazard to operators, and the discharge of experimental waste liquid does not conform to the current environmental protection concept

Method used

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  • A kind of method and kit for detecting dha content in blood sample
  • A kind of method and kit for detecting dha content in blood sample

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Experimental program
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Embodiment

[0032] (1) Blood sample pretreatment

[0033] Add 1ml acetonitrile / 37% hydrochloric acid (4:1, v / v) solution to 200μl serum sample, incubate at 90°C for 2h after vortexing, then cool to room temperature, add 2ml hexane and vortex for 20s, room temperature Let stand for 5 minutes and then centrifuge at 3000 rpm for 1 minute. Take the uppermost part of the solution into a sample bottle, dry it with nitrogen, redissolve the sample in 200 μl methanol aqueous solution (180:20, v / v), filter through a 0.2 μm filter membrane, and obtain the sample to be tested;

[0034] (2) Preparation of standard working solution: prepare 100 μg / mL standard stock solution with water, and then use 5% bovine serum albumin methanol solution (m / v) to prepare 7 gradients, respectively 100, 200, 500, 1000, 5000, 10000, 20000, 50000ng / ml. The standard solution was divided into 1.5mL brown bottles and stored at -20°C for later use;

[0035] (3) HPLC-MS / MS detection

[0036] Chromatographic column: C18 co...

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Abstract

The invention belongs to the field of detection of unsaturated fatty acid and in particular relates to a method for detecting DHA (Docosahexaenoic Acid) content in a blood sample and kit. According tothe method, the DHA content in the blood sample is detected by adopting HPLC-MS / MS (High Performance Liquid Chromatography-Mass Spectrometry / Mass Spectrometry); the method comprises the following steps: (1) pre-treating a blood sample; (2) preparing a standard working solution; (3) carrying out HPLC-MS / MS detection. By adopting the method, the aim of accurately detecting DNA in a blood serum sample by applying an HPLC-MS / MS technology is realized for the first time and a matrix effect is reduced; the method is simple, convenient and rapid to operate, high in flux and low in cost, is used foreffectively monitoring a DHA level in a human body and has guidance meaning on reasonable and safe supplementation of DHA; the method is easy for clinical popularization and generalization.

Description

technical field [0001] The invention belongs to the detection field of unsaturated fatty acids, and in particular relates to a method and a kit for detecting DHA content in blood samples. Background technique [0002] DHA (Docosahexaenoic acid, 22:6△4.7.10.13.16.19, full name docosahexaenoic acid) is an important long-chain polyunsaturated fatty acid (PUFA for short), which is the main component of human brain and neurons. Major lipid component in retinal cell membranes. In the cells of the rod-shaped outer part of the retina, DHA can reach more than 60% of the total lipids in the cells, and in the cells of the human brain tissue, DHA accounts for about 10% of the total lipids. However, humans cannot synthesize DHA by themselves and must be ingested through food. During pregnancy, if a woman lacks DHA in her diet, it will lead to insufficient DHA supplementation for the fetus, resulting in abnormal growth and development of brain cells, resulting in mental retardation; sev...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 冯振景叶松弭兆元
Owner JINAN YING SHENG BIOTECH